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Molecular profile of non-aflatoxigenic phenotype in native strains of Aspergillus flavus

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Abstract

Aflatoxins are the most common mycotoxin contaminant reported in food and feed. Aflatoxin B1, the most toxic among different aflatoxins, is known to cause hepatocellular carcinoma in animals. Aspergillus flavus and A. parasiticus are the main producers of aflatoxins and are widely distributed in tropical countries. Even though several robust strategies have been in use to control aflatoxin contamination, the control at the pre-harvest level is primitive and incompetent. Therefore, the aim of the study was to isolate and identify the non-aflatoxigenic A. flavus and to delineate the molecular mechanism for the loss of aflatoxin production by the non-aflatoxigenic isolates. Eighteen non-aflatoxigenic strains were isolated from various biological sources using cultural and analytical methods. Among the 18 isolates, 8 isolates produced sclerotia and 17 isolates had type I deletion in norB-cypA region. The isolates were confirmed as A. flavus using gene-specific PCR and sequencing of the ITS region. Later, aflatoxin gene-specific PCR revealed that the defect in one or more genes has led to non-aflatoxigenic phenotype. The strain R9 had maximum defect, and genes avnA and verB had the highest frequency of defect among the non-aflatoxigenic strains. Further, qRT-PCR confirmed that the non-aflatoxigenic strains had high frequency of defect or downregulation in the late pathway genes compared to early pathway genes. Thus, these non-aflatoxigenic strains can be the potential candidates for an effective and proficient strategy for the control of pre-harvest aflatoxin contamination.

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Acknowledgements

The authors are thankful to Director, CSIR- Central Food Technological Research Institute, for providing necessary facilities to carry out this study and CSIR for the financial support in the form of Junior and senior research fellowship to the first author. Authors would also like to thank Dr. Anu Appaiah K. A., Principal scientist, Microbiology and Fermentation Technology, CSIR-CFTRI, for his kind support.

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Authors and Affiliations

  1. Microbiology and Fermentation Technology, CSIR-Central Food Technological Research Institute, Mysuru, Karnataka, 570 020, India

    K. Raksha Rao, A. V. Vipin & G. Venkateswaran

  2. Academy of Scientific and Innovative Research (AcSIR), CSIR-Central Food Technological Research Institute Campus, Mysuru, Karnataka, 570 020, India

    K. Raksha Rao, A. V. Vipin & G. Venkateswaran

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  1. K. Raksha Rao
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Correspondence to G. Venkateswaran.

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203_2020_1822_MOESM1_ESM.docx

Fig.S1. Screening and isolation Aspergillus flavus from soil and seed samples. a. Screening through standard plate method; b. Screening through standard blotter method; c.Aspergillus flavus on PDA; d. Microscopic view of A. flavus; e. Culture methods to differentiate aflatoxigenic and non-aflatoxigenic A. flavusi. Ultraviolet test; ii. Ammonia vapor test; iii. Yellow pigmentation (DOCX 1015 kb)

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Rao, K.R., Vipin, A.V. & Venkateswaran, G. Molecular profile of non-aflatoxigenic phenotype in native strains of Aspergillus flavus. Arch Microbiol 202, 1143–1155 (2020). https://doi.org/10.1007/s00203-020-01822-1

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